The practical proceeded in five sessions. The first session involved DNA manipulation, which involved two steps. The aim of the first step, small-scale plasmid isolation (â€˜Miniprepâ€™), was to isolate a recipient plasmid, pBlue Script KS II (+), from anÂ E. coliÂ culture prepared the previous day. In the second exercise, a double restriction endonuclease digestion of the two plasmids, i.e., the donor (pProEX) and the recipient (pBKSII), was done usingÂ BamHI andÂ HindIII. The aims of this step were to excise the gene of interest (Pc-FtsZ) from pProEX and prepare the recipient plasmid, pBKSII, for cloning. The two restriction endonucleases cleaved the DNA at specific sites to generate sticky ends that allowed the foreign fragment (Pc-FtsZ) to be inserted into the recipient plasmid.
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